Avian Antibodies for Staphylococcal Enterotoxin B as an Efficient Tool for FRET-Based Fluoroimmunosensor

Vinayaka, A. C. and Muthukumar, S. P. and Thakur, M. S. (2013) Avian Antibodies for Staphylococcal Enterotoxin B as an Efficient Tool for FRET-Based Fluoroimmunosensor. BioNanoScience, 3. pp. 232-240.

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Abstract

In recent years, antibodies have become an indispensable tool in research, diagnostic, therapy and other laboratory applications. In this direction, hens have become an alternate source for polyclonal antibody production necessary for immunosensing applications. Herein, we present generation of avian antibody (IgY) against staphylococcal enterotoxin B (SEB) used as an efficient bio-recognition molecule while developing Forster resonance energy transfer (FRET)-based fluorescence inhibition assay format for monitoring SEB, a bacterial super-antigen responsible for food poisoning. Immunization of white leghorn hens with SEB, generated antibodies (IgY) with highest yield of 51.1 mg in hyperimmune egg having affinity constant of 1.81×1010 M−1 that was bioconjugated with gold nanoparticle (GNP) by electrostatic interaction. Quantum dot (QD, CdTe523) and GNP bioconjugated to SEB and anti-SEB IgY, respectively, were used as donor–acceptor pair during FRET to study resonance interactions and fluorescence quenching in an immunocomplex. The mutual affinity between IgY-GNP and SEB-CdTe523 was made use to obtain efficient energy transfer between respective chromophores. The presence of SEB in the range 1,000 to 20 ng mL−1 varied the rate of fluorescence quenching due to FRET in SEB-CdTe523 demonstrating efficiency of fluorescence inhibition assay. It was possible to achieve a detection limit of 8.53 ng mL−1 with a regression coefficient R2=0.9813. Thus, bioconjugation of nanoparticles demonstrated their efficiency in sensitive monitoring of SEB through immuno-affinity reactions to address the potential health risk and economic impacts of staphylococcal food poisoning.

Item Type:	Article
Uncontrolled Keywords:	Avian antibodies (IgY) . Enterotoxins . Food poisoning . FRET . Immunosensing . Quantum dots
Subjects:	600 Technology > 08 Food technology > 12 Allergy & Toxicology > 02 Poisoning
Divisions:	Dept. of Biochemistry Fermentation Technology and Bioengineering
Depositing User:	Food Sci. & Technol. Information Services
Date Deposited:	23 Sep 2016 11:28
Last Modified:	23 Sep 2016 11:28
URI:	http://ir.cftri.res.in/id/eprint/12325

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